Patent Application
20230322723
This application claims priority to Korean Patent Application No. 10-2020-0117186 filed on Sep. 11, 2020, and all contents disclosed in the specification and drawings of the application are incorporated in this application by reference.
The present disclosure relates to a group of compounds for inhibiting, i.e. treating or preventing cancers exhibiting EGFR mutations. The present disclosure also relates to pharmaceutical compositions comprising such compound. The present disclosure also relates to useful methods for treating or preventing cancers exhibiting EGFR mutations, in particular, cancers exhibiting EGFR mutations of any one or more of del19, T790M, C797S, and L858R, using such compound. That is, the present disclosure relates to the medical use of the compounds according to the present invention for treating or preventing said cancer.
Epidermal growth factor receptor (EGFR) is a protein composed of a receptor part and a tyrosine kinase part, and serves to transmit signals from outside the cell to the inside of the cell through the cell membrane. EGFR plays an essential role in normal cell regulation through intracellular signal transduction. However, overexpression of EGFR or activating EGFR mutations characterized by ligand-independent tyrosine kinase activity are known to induce growth, differentiation, angiogenesis, metastasis, resistance expression, etc. of cancer cells by abnormally activating the cell signaling system. It has been reported that EGFR is abnormally overexpressed or frequently mutated in most solid cancer cells, which is associated with poor prognosis. For example, lung cancer, liver cancer, esophageal cancer, stomach cancer, colon cancer, small intestine cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, parathyroid cancer, kidney cancer, cervical cancer, sarcoma, prostate cancer, urethral cancer, bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, psoriasis, and fibroadenoma are known to be associated with EGFR mutations (Cancer Communications. 2020, 40:43-59).
Among them, EGFR activating mutations such as the L858R point mutation in exon 21 of the EGFR tyrosine kinase domain or the in-frame deletion in exon 19 are known to be important causes of non-small cell lung cancer. Therefore, according to the prediction that the anticancer effect will be excellent if cancer cell signaling through the epidermal growth factor receptor is blocked, research for developing anticancer drugs targeting the epidermal growth factor receptor is being actively conducted.
The first drug developed as an EGFR tyrosine kinase inhibitor among small molecules is Gefitinib, which is a reversible inhibitor that selectively inhibits EGFR (Erb-B1) among EGFR subtypes. Another drug with such characteristics is Erlotinib, and this EGFR targeting therapy is mainly used for patients with EGFR activating mutations for non-small cell lung cancer (NSCLC) as a main indication.
However, it has been reported that NSCLC patients with EGFR activating mutations treated with Gefitinib or Erlotinib develop resistance to the drug after about 8 to 16 months, and about 60% of them show resistance due to the EGFR T790M mutation. (Helena A. Yu et al., Clin Cancer Res. 19(8), 2240, 2013).
Irreversible inhibitors have been proposed to overcome resistance to existing EGFR inhibitors such as Gefitinib or Erlotinib. However, since EGFR irreversible inhibitors also have high activity against EGFR WT (wild-type), which also exists in normal cells, serious side effects are caused when doses to overcome resistance due to EGFR T790M mutation are administered. It indicates the limitations of clinical application.
As an alternative to this, a number of drugs including Osimertinib, Olmutinib, Naquotinib, and Avitinib, which are EGFR mutation-selective inhibitors, are under development in the clinical stage. However, according to the clinical results of Osimertinib for non-small cell lung cancer patients with EGFR-resistant mutations, after about 10 months, other resistance mechanisms are activated, resulting in drug resistance, among which the C797S mutation appears at a high rate of 20% or more. The C797S mutation is a point mutation in which cysteine 773 (Cys773), which forms a covalent bond with irreversible inhibitors of EGFR, is changed to serine. As a result, it is impossible to form a covalent bond with irreversible inhibitors of EGFR, thereby causing a decrease in reactivity to the drug.
As described above, the development of EGFR-targeted therapeutics shows limitations that cannot maintain drug efficacy over a certain period of time due to the expression of primary and secondary resistance. In particular, researches on the EGFR C797S mutation have not presented even a substance under clinical study other than reports on early-stage preclinical studies, so effective treatment for this is urgently required.
Therefore, a problem to be solved by the present disclosure is to provide a compound having an activity suppressing, inhibiting and/or degrading EGFR mutation-expressing cancer, pharmaceutical compositions comprising the compound as an active ingredient, and its medical use for treating or preventing cancer showing EGFR mutation(s).
Another problem to be solved by the present disclosure is to provide a method for treating or alleviating EGFR mutation-expressing cancer, comprising administering to a patient in need of treatment, improvement or prevention of EGFR mutation-expressing cancer the compound according to the present invention, wherein it inhibits EGFR mutation-expressing cancer cells.
In order to solve the above problem, one embodiment of the present invention provides a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof.
In the Chemical Formula 1,
Compounds of the present disclosure are excellent in degrading EGFR mutant proteins in cancers with EGFR mutations. Therefore, a preventive or therapeutic effect on cancers exhibiting EGFR mutations can be expected.
In particular, the compounds of the present disclosure have the advantage of differentially degrading mutant EGFR proteins including C797S and the like rather than EGFR proteins of cells having normal EGFR.
The compounds according to the present disclosure are particularly useful for degradation of EGFR mutant proteins exhibiting any one or more mutations among Del19, T790M, C797S, and L858R, and are particularly useful for degradation of Del19/T790M/C797S and L858R/T790M/C797S mutant proteins. Mutant EGFR including C797S is a resistance mutation that appears after treatment with Tagrisso (Osimertinib), a third-generation therapeutic agent. The compounds of the present disclosure have the advantage of being useful for inhibiting, degrading, and the like in lung cancer having the C797S mutation of EGFR, in particular, non-small cell lung cancer (NSCLC) cells.
In the compounds of the present disclosure, the right moiety in the chemical structure serves to bind and/or inhibit mutant EGFR protein, etc., and the left moiety serves as an E3 ligase binder to induce ubiquitin-proteasome system (UPS). This is expected to further promote degradation and/or inhibition of the protein, but the present invention is not limited to these theoretically predicted mechanisms.
In another embodiment, the present disclosure provides a pharmaceutical composition comprising a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or additive.
In the other embodiment, the present disclosure provides a method for treating cancer exhibiting an EGFR mutation, comprising administering to a subject a therapeutically effective amount of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof. Cancer exhibiting the EGFR mutation is, for example, lung cancer, liver cancer, esophageal cancer, stomach cancer, colon cancer, small intestine cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, parathyroid cancer, kidney cancer, cervical cancer, sarcoma, prostate cancer, urethral cancer, bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, psoriasis, fibroadenoma, and the like. Preferably, the cancer exhibiting an EGFR mutation is lung cancer. More preferably, the cancer exhibiting an EGFR mutation is non-small cell lung cancer.
That is, the present disclosure provides a medical use characterized by using the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof according to the present invention as an active ingredient. In one embodiment, the medical use of the present invention is for the treatment or prevention of a disease or condition described herein.
The compounds, pharmaceutical compositions comprising such compounds, and their medical uses are more fully described in the detailed description that follows.
The following description is illustrative only and is not intended to limit the invention, application, or use.
The following terms used herein are defined as follows.
As used herein, the terms “substituent”, “radical”, “group”, “moiety”, and “fragment” may be used interchangeably.
As used herein, the term “alkyl” means a saturated straight chain or branched non-cyclic hydrocarbon, unless the context clearly dictates otherwise, having from 1 to 10 carbon atoms. “Lower alkyl” means alkyl having from 1 to 4 carbon atoms. Representative saturated straight chain alkyls include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, -n-octyl, -n-nonyl and -n-decyl, while saturated branched alkyls include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimethylpentyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl, 3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like.
As used herein, if the term “C1-6”, “C1-6”, or “C1-C6” is used, it means the number of carbon atoms is from 1 to 6. For example, C1-6alkyl means an alkyl which carbon number is any integer of from 1 to 6.
As used herein, the terms “halogen” and “halo” mean fluorine, chlorine, bromine or iodine. In a preferred embodiment of the present invention, the halogen is chlorine or fluorine.
As used herein, the term “haloalkyl”, “haloalkoxy”, “haloalkenyl”, or “haloalkynyl” mean an alkyl, alkoxy, alkenyl or alkynyl group, wherein one or more hydrogen atoms are substituted with halogen atoms. For example, the haloalkyl includes —CF3, —CHF2, —CH2F, —CBr3, —CHBr2, —CH2Br, —CC13, —CHC12, —CH2Cl, —Cl3, —CHI2, —CH2I, —CH2—CF3, —CH2—CHF2, —CH2—CH2F, —CH2—CBr3, —CH2—CHBr2, —CH2—CH2Br, —CH2—CC13, —CH2—CHC12, —CH2—CH2Cl, —CH2—Cl3, —CH2—CHI2, —CH2—CH2I, and the like, wherein alkyl and halogen are as described above. In an embodiment of the present invention, haloalkyl is —CF3.
As used herein, the term “alkoxy” means —O-(alkyl) including —OCH3, —OCH2CH3, —O(CH2)2CH3, —O(CH2)3CH3, —O(CH2)4CH3, —O(CH2)5CH3, and the like, wherein alkyl is as defined above. As used herein, the term “lower alkoxy” means —O-(lower alkyl), wherein lower alkyl is as defined above.
The terms “aryloxy”, “cycloalkyloxy” or “heterocycloalkyloxy” are RO—, where R is aryl, cycloalkyl or heterocycloalkyl, respectively, as defined herein. “Arylthio” is RS—, where R is aryl as defined above.
As used herein, the term “aryl” means a carbocyclic aromatic group containing from 3 to 10 ring atoms. Representative examples include, but are not limited to, phenyl, tolyl, xylyl, naphthyl, tetrahydronaphthyl, anthracenyl, fluorenyl, indenyl, and azulenyl. A carbocyclic aromatic group can be unsubstituted or optionally substituted.
As used herein, the term “cycloalkyl” means a monocyclic or polycyclic saturated ring having carbon and hydrogen atoms and having no carbon-carbon multiple bonds. Examples of monocyclic rings include, but are not limited to, (C3-C7)cycloalkyl groups, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Examples of polycyclic rings include, but are not limited to, fused bicyclic rings (bicyclic hydrocarbon rings) such as octahydropentalene and decahydronaphthalene; spiro rings such as spiro[3.3]heptane, spiro[3.4]octane, spiro[3.5]nonane, spiro[4.4]nonane, spiro[4.5]decane, and spiro[5.5]undecane; and bridged bicycle rings such as bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, and bicyclo[2.2.2]octane. A cycloalkyl group can be unsubstituted or optimally substituted. In an embodiment of the present invention, the cycloalkyl group is monocyclic ring. In an embodiment of the present invention, cycloalkyl is cyclopropyl. In an embodiment of the present invention, “spiro hydrocarbon ring” or “spiro ring” is spiro[3.3]heptane, spiro[3.4]octane, spiro[4.3]octane, spiro[4.5]decane, spiro[4.4]decane, or spiro[5.5]undecane.
As used herein, the term “heteroaryl” means an aromatic heterocycle ring of 3- to 10-members and having at least one heteroatom selected from nitrogen, oxygen and sulfur, and containing at least 1 carbon atom, including both mono- and bicyclic ring systems. Representative heteroaryls are furan, 4H-pyran, pyrrole, imidazole, pyrazole, triazole, tetrazole, pyridine, pyrimidine, pyridazine, pyrazine, triazine, thiophene, ozaxole, isoxazole, thiazole, isothiazole, oxadiazole, benzofuran, benzothiophene, quinoline, indole, benzoxazole, benzimidazole, benzothiazole, cinnoline, phthalazine, quinazoline, 1H-azepine and the like.
The term “heterocycle” or “heterocycloalkyl” means a 3- to 7-membered monocyclic, or 6- to 12-membered bicyclic, saturated heterocyclic ring which contains from 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, and wherein the nitrogen and sulfur heteroatoms can be optionally oxidized, and the nitrogen heteroatom can be optionally quaternized. Representative heterocycles include oxiran, oxetan, tetrahydrofuran, tetrahydropyran, 1,4-dioxane, aziridine, azetidine, pyrrolidine, piperidine, piperazine, pyrrolidinone, hydantoine, valerolactam, thiirane, thietane, tetrahydrothiophene, tetrahydrothiopyra, morpholine, tetrahydropyridine, and tetrahydropyrimidine. Heterocycles include a bicyclic ring in which part of the heterocycle is fused to a benzene or cyclopenta-1,3-diene ring. The heterocycle can be attached via any heteroatom or carbon atom. In addition, heterocycles include fused bicyclic rings, spiro rings and bridged bicyclic rings in which one or more carbon atoms of the aforementioned polycyclic rings are replaced with nitrogen, oxygen or sulfur atoms. For example, when the heteroatom is nitrogen, these include, but are limited to, fused heterobicyclic rings (bicyclic hetero rings) such as octahydrocyclopenta[c]pyrrole, octahydropyrrolo[3,4-c]pyrrole, decahydroisoquinoline, and decahydro-2,6-naphthyridine; spiro (hetero)rings such as 2-azaspiro[3.3]heptane, 2,6-diazaspiro[3.3]heptane, 2-azaspiro[3.4]octane, 2,6-diazaspiro[3.4]octane, 2-azaspiro[3.5]nonane, 2,7-diazaspiro[3.5]nonane, 2-azaspiro[4.4]nonane, 2,7-diazaspiro[4.4]nonane, 8-azaspiro[4.5]decane, 2,8-diazaspiro[4.5]decane, 3-azaspiro[5.5]undecane, and 3,9-diazaspiro[5.5]undecane; and bridged heterobicyclic rings such as 2-azabicyclo[2.1.1]hexane, 2-azabicyclo[2.2.1]heptane, 2,5-diazabicyclo[2.2.1]heptane, 2-azabicyclo[2.2.2]octane, and 2,5-diazabicyclo[2.2.2]octane. In an embodiment of the present invention, the spiro heteroring is 2-azaspiro[3.3]heptane, 2,6-diazaspiro[3.3]heptane, 3-azaspiro[5.5]undecane, 3,9-diazaspiro[5.5]undecane, or 1,4-dioxaspiro[4.5]decane.
“Heterocycle fused to a phenyl” means a heterocycle attached to two adjacent carbon atoms of a phenyl ring, wherein the heterocycle is as defined above.
In this specification, “*” or “” means connected to another moiety.
As used herein, the term “pharmaceutically acceptable salt(s)” refers to a salt prepared from active compounds according to the present disclosure with relatively non-toxic acids or bases, depending on the particular substituents of those compounds. When the compounds have a relatively acidic group, base-added salts can be obtained by contacting the neutral compounds with a sufficient amount of the desired base and a pure or inert solvent. Suitable pharmaceutically acceptable base addition salts include, but are not limited to sodium, potassium, calcium, aluminum, organic amino, magnesium salts and the like. When the compounds have a relatively basic group, acid-added salts can be obtained by contacting the neutral compounds with a sufficient amount of the desired acid and pure or inert solvent. Suitable pharmaceutically acceptable acid addition salts include salts derived from non-toxic organic acids including, but are not limited to, acetic acid, propionic acid, isobutyl acid, oxalic acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like, and non-toxic inorganic acids including, but are not limited to, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydrogen iodide, phosphorous acid and the like. Also it includes a salt of amino acid such as arginate or its analogues, and it also includes analogues of organic acid such as glucuronic or galacturonic acid. Some specific compounds of this disclosure have both basic and acidic functionality for the conversion of compounds with a basic or acidic portion (addition) salts. Other examples of salts are well known through literature known in the art to which the present invention pertains.
As used herein, “effective amount” refers to an amount of a compound of the invention sufficient to slow or minimize the progression of a cancer with EGFR mutation or to provide a therapeutic benefit in the treatment or management of a cancer with EGFR mutation. “Effective amount” also refers to an amount sufficient to inhibit or reduce EGFR mutation proteins, either in vitro or in vivo.
As used herein, the term “treatment” may be any one or more of preventive treatment, palliative treatment, and/or restorative treatment.
As used herein, the phrase “compound(s) of this/the invention” includes any compound(s) of Chemical Formula 1, as well as clathrates, hydrates, solvates, or polymorphs thereof. And, even if the term “compound(s) of the invention” does not mention its pharmaceutically acceptable sat, the term includes salts thereof. In one embodiment, the compounds of this disclosure include stereo-chemically pure compounds, e.g., those substantially free (e.g., greater than 85% ee, greater than 90% ee, greater than 95% ee, greater than 97% ee, or greater than 99% ee) of other stereoisomers. That is, if the compounds of Chemical Formula 1 according to the present disclosure or salts thereof are tautomeric isomers and/or stereoisomers (e.g., geometrical isomers and conformational isomers), such isolated isomers and their mixtures also are included in the scope of this disclosure. If the compounds of the present disclosure or salts thereof have an asymmetric carbon in their structures, their active optical isomers and their racemic mixtures also are included in the scope of this disclosure.
As used herein, the term “polymorph” refers to solid crystalline forms of a compound of this disclosure or complex thereof. Different polymorphs of the same compound can exhibit different physical, chemical and/or spectroscopic properties. Different physical properties include, but are not limited to stability (e.g., to heat or light), compressibility and density (important in formulation and product manufacturing), and dissolution rates (which can affect bioavailability). Differences in stability can result from changes in chemical reactivity (e.g., differential oxidation, such that a dosage form discolors more rapidly when comprised of one polymorph than when comprised of another polymorph) or mechanical characteristics (e.g., tablets crumble on storage as a kinetically favored polymorph converts to thermodynamically more stable polymorph) or both (e.g., tablets of one polymorph are more susceptible to breakdown at high humidity). Different physical properties of polymorphs can affect their processing. For example, one polymorph might be more likely to form solvates or might be more difficult to filter or wash free of impurities than another due to, for example, the shape or size distribution of particles of it.
As used herein, the term “solvate” means a compound or its salt according to this disclosure that further includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Preferred solvents are volatile, non-toxic, and acceptable for administration to humans in trace amounts.
As used herein, the term “hydrate” means a compound or its salt according to this disclosure that further includes a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.
As used herein, the term “clathrate” means a compound or its salt in the form of a crystal lattice that contains spaces (e.g., channels) that have a guest molecule (e.g., a solvent or water) trapped within.
As used herein, the term “purified” means that when isolated, the isolate is greater than 90% pure, in one embodiment greater than 95% pure, in another embodiment greater than 99% pure and in another embodiment greater than 99.9% pure.
The term “hydrido” refers to a single —H atom (H) and is used interchangeably with the symbol “H” or the term “hydrogen”.
If a substituent is described as “optionally substituted”, the substituent may be (1) unsubstituted or (2) substituted with one or more of the defined substituents. If the substitutable position is unsubstituted, the default substituent is hydrogen.
As used herein, the singular “a” and “an” may include the plural forms unless the context clearly dictates otherwise.
The term “pharmaceutically acceptable” means suitable for use as a pharmaceutical preparation, and generally considered safe for such use. The term also means that it has been officially approved by the governing body of a country for this use, or is listed in the Korean Pharmacopoeia or the United States Pharmacopoeia.
Compound of the Present Invention
One embodiment of the present invention provides a compound having a structure of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.
In the Chemical Formula 1,
In a preferred embodiment, the present invention provides a compound represented by the Chemical Formula 1 or a pharmaceutically acceptable salt thereof, wherein
R11 is hydrogen, C1˜C6 alkyl, or (CH2)1-6—C(O),
In the other embodiment, the present invention provides a compound represented by the Chemical Formula 1 or a pharmaceutically acceptable salt thereof, wherein L is a direct bond or divalent, saturated or unsaturated C1˜C50 hydrocarbon chain; wherein at least one of the methylene units of L may be replaced with one or more selected from the group consisting of -Cy-, —O—, —N(R20)—, —S—, —OC(O)—, —C(O)O—, —C(O)—, —S(O)—, —S(O)2—, —N(R20)S(O)2—, —S(O)2N(R20)—, —N(R20)C(O)—, —C(O)N(R20)—, —OC(O)N(R20)—, —N(R20)C(O)—, —(CH2OCH2)—, —(OCH2CH2)—, —(CH2CH2O)—, —(C(O)CH2CH2)—, and —(CH2CH2C(O))—; here, -Cy- is each independently a divalent ring substituent, which is phenylene, 4-7-membered spiro hydrocarbon ring, 8-10-membered bicyclic hydrocarbon ring, 4-7-membered heterocyclic ring, 4-7-membered spiro heterocyclic ring, 8-10-membered bicyclic heterocyclic ring, 5-6-membered heteroarylene, or 8-10-membered bicyclic heteroarylene, wherein R20 is hydrogen or C1˜C4 alkyl.
Preferably, in the other embodiment, the present invention provides a compound represented by the Chemical Formula 1 or a pharmaceutically acceptable salt thereof, wherein the X-L of the Chemical Formula 1 is —K1-K2-K3-K4-K5-,
More preferably, another embodiment of the present invention provides a compound or a pharmaceutically acceptable salt thereof, wherein X-L of the Chemical Formula 1 is the X-L moiety used in the following examples. X-L used in the examples may be connected to various Bs (except for X) disclosed herein on the left side, and various ‘-A-EGFR-binding moieties’ disclosed herein may be connected to the right side.
More preferably, in the other embodiment, the present invention provides a compound represented by the Chemical Formula 1 or a pharmaceutically acceptable salt thereof, wherein the A is
Preferably, one embodiment of the present invention provides a compound represented by the Chemical Formula 1a below or a pharmaceutically acceptable salt thereof
In the Chemical Formula 1a,
Among the compounds of Chemical Formula 1, when they have the same structure as Chemical Formula 1a, bioavailability is significantly improved as described later. As shown in the exemplary experimental results described later, Compounds 74, 76, 77, 80 and 81 showed significantly improved AUC compared to the corresponding compounds 45, 71, 51, 65 and 53.
Preferably, one embodiment of the present invention also provides a compound represented by the Chemical Formula 1 or Chemical Formula 1a or a pharmaceutically acceptable salt thereof, wherein X is O—(CH2)0-6, N(R13)—(CHR14), or O—(CHR14), and R14 is C1˜C6 alkyl (preferably, methyl) (the rest of the substituents are the same as defined in Chemical Formula 1 or 1a). As shown in the exemplary experimental results described later, Compounds 69, 70, 75, 78, and 82 showed significantly improved AUC compared to the corresponding compounds 53 and 65. These compounds are compounds in which K1 of —K1-K2-K3-K4-K5- is *—O—C1˜C6 alkyl-*, or *—NH—C1˜C6 alkyl-*(K2, K3, K4, and K5 are the same as the previous definition), wherein O or NH is connected to the 2nd or 3rd carbon other than the terminal of C1˜C6 alkyl.
The present inventors performed various evaluation experiments after synthesizing various compounds in order to secure compounds with good therapeutic or preventive effects for cancer showing EGFR mutations and reduced other side effects and their use by having particularly high inhibitory, degrading and/or binding activity to the mutant EGFR protein, and also high selectivity thereto, and preferably effectively degrading the mutant EGFR protein. The present inventors also evaluated pharmacokinetic aspects such as bioavailability in addition to pharmacological aspects. Finally, the present invention was completed by confirming that the compounds of the present invention are suitable for the purpose of the present invention.
One embodiment of the present invention provides the compounds of Examples described below and pharmaceutically acceptable salts thereof as non-limiting examples of the compounds of Chemical Formula 1 according to the present invention.
Preferably, another embodiment of the present invention provides a compound and a pharmaceutically acceptable salt thereof, wherein the compound of Chemical Formula 1 is a compound of Table 1 below. The compounds of Table 1 below are more preferable in various aspects such as drug efficacy and bioavailability.
In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof according to the present invention, and a pharmaceutically acceptable carrier.
Medical Uses and Methods of Treatment of the Compounds According to the Present Invention
The present invention further provides methods for treating a disease or condition in a subject having or susceptible to having such a disease or condition, by administering to the subject a therapeutically-effective amount of one or more compounds as described above. In one embodiment, the treatment is preventative treatment. In another embodiment, the treatment is palliative treatment. In another embodiment, the treatment is restorative treatment.
1. Diseases or Conditions
The compounds of the present invention for inhibiting or/and degrading EGFR mutant protein are useful for various therapeutic or prophylactic uses. These compounds can be used to suppress or inhibit EGFR mutant protein activity, and can also be used to treat cancers showing an EGFR mutant or to prevent exacerbation of these diseases. Accordingly, the present invention provides a method of inhibiting, suppressing, and/or degrading EGFR mutant protein activity in a cell. In this method, the cells are contacted with an effective amount of a compound of the present invention. In one embodiment, the cell is in a subject (e.g., a lung cancer patient). The method of the present invention comprises administering a pharmaceutical composition comprising a therapeutically or prophylactically effective amount of a compound of the present invention to a subject in need of treatment or prevention.
In one embodiment, the present invention provides a method for inhibiting, degrading or inhibiting EGFR mutant protein activity in cancer cells exhibiting EGFR mutations. For example, the present invention can be used for treatment or prevention of lung cancer, liver cancer, esophageal cancer, gastric cancer, colon cancer, small intestine cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, parathyroid cancer, kidney cancer, cervical cancer, sarcoma, prostate cancer, urethral cancer, bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, psoriasis, fibroadenoma, and the like, which have EGFR mutations.
A compound of the present invention can be administered to the subject in the form of a pharmaceutical composition described herein.
In another embodiment, the present invention provides a method for treating or preventing cancer exhibiting an EGFR mutation in a subject, and cancers exhibiting such an EGFR mutation include lung cancer, liver cancer, esophageal cancer, gastric cancer, colon cancer, small intestine cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, parathyroid cancer, kidney cancer, cervical cancer, sarcoma, prostate cancer, urethral cancer, bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, psoriasis, fibroadenoma, and the like. Such methods comprise administering to a subject in need of treatment an amount of a compound of the present invention sufficient to inhibit EGFR mutant protein activity, i.e., a therapeutically effective amount.
In particular, the compounds of the present invention are useful for degradation of EGFR mutant proteins exhibiting any one or more mutations among Del19, T790M, C797S, and L858R, and are particularly useful for degradation of Del19/T790M/C797S and L858R/T790M/C797S EGFR mutant proteins. Cancers with these mutant EGFRs may be lung cancers, particularly non-small cell lung cancer (NSCLC).
2. Subjects
Suitable subjects to be treated according to the present invention include mammalian subjects. Mammals according to the present disclosure include, but are not limited to, human, canine, feline, bovine, caprine, equine, ovine, porcine, rodents, lagomorphs, primates, and the like, and encompass mammals in utero. Subjects can be of both sexes and can be at any stage of development.
In one embodiment, the suitable subject to be treated according to the present invention is human.
3. Administration and Dosing
The compounds of the present invention are generally administered in a therapeutically effective amount.
The compounds of the present invention can be administered by any suitable route in the form of a pharmaceutical composition adapted to such a route, and in a dose effective for the treatment intended. An effective dosage is typically in the range of about 0.001 to about 100 mg per kg body weight per day, preferably about 0.01 to about 50 mg/kg/day, in single or divided doses. Depending on age, species and disease or condition being treated, dosage levels below the lower limit of this range may be suitable. In other cases, still larger doses may be used without harmful side effects. Larger doses may also be divided into several smaller doses, for administration throughout the day.
Pharmaceutical Compositions, Dosage Forms and Administration Routes
For the treatment of the diseases or conditions referred to above, the compounds described herein or pharmaceutically acceptable salts thereof can be administered as follows:
Oral Administration
The compounds of the present invention may be administered orally, including by swallowing, so that the compound enters the gastrointestinal tract, or absorbed into the blood stream directly from the mouth (e.g., buccal or sublingual administration).
Suitable compositions for oral administration include solid, liquid, gel or powder formulations, and have a dosage form such as tablet, lozenge, capsule, granule or powder.
Compositions for oral administration may optionally be enteric coated and may exhibit delayed or sustained release through the enteric coating. That is, the composition for oral administration according to the present invention may be a formulation having an immediate or modified release pattern.
Liquid formulations can include solutions, syrups and suspensions, which can be used in soft or hard capsules. Such formulations may include a pharmaceutically acceptable carrier, for example, water, ethanol, polyethylene glycol, cellulose, or an oil. The formulation may also include one or more emulsifying agents and/or suspending agents.
In a tablet dosage form the amount of drug, active ingredient, present may be from about 0.05% to about 95% by weight, more typically from about 2% to about 50% by weight of the dosage form. In addition, tablets may contain a disintegrant, comprising from about 0.5% to about 35% by weight, more typically from about 2% to about 25% of the dosage form. Examples of disintegrants include, but are not limited to, lactose, starch, sodium starch glycolate, crospovidone, croscarmellose sodium, maltodextrin, or mixtures thereof.
Suitable lubricants, for use in a tablet, may be present in amounts from about 0.1% to about 5% by weight, and include, but are not limited to, talc, silicon dioxide, stearic acid, calcium, zinc or magnesium stearate, sodium stearyl fumarate and the like.
Suitable binders, for use in a tablet, include, but are not limited to, gelatin, polyethylene glycol, sugars, gums, starch, polyvinyl pyrrolidone, hydroxypropyl cellulose, hydroxypropylmethyl cellulose and the like. Suitable diluents, for use in a tablet, include, but are not limited to, mannitol, xylitol, lactose, dextrose, sucrose, sorbitol, microcrystalline cellulose and starch.
Suitable solubilizers, for use in a tablet, may be present in amounts from about 0.1% to about 3% by weight, and include, but are not limited to, polysorbates, sodium lauryl sulfate, sodium dodecyl sulfate, propylene carbonate, diethyleneglycol monoethyl ether, dimethyl isosorbide, polyethylene glycol (natural or hydrogenated) castor oil, HCOR™ (Nikkol), oleyl ester, Gelucire™, caprylic/caprylic acid mono/diglyceride, sorbitan fatty acid esters, and Solutol HS™.
Parenteral Administration
Compounds of the present disclosure may be administered directly into the blood stream, muscle, or internal organs. Suitable means for parenteral administration include intravenous, intra-muscular, subcutaneous intraarterial, intraperitoneal, intrathecal, intracranial, and the like. Suitable devices for parenteral administration include injectors (including needle and needle-free injectors) and infusion methods.
Compositions for parenteral administration may be formulated as immediate or modified release, including delayed or sustained release.
Most parenteral formulations are liquid compositions, and the liquid composition is an aqueous solution containing the active ingredient according to the present invention, a salt, a buffering agent, an isotonic agent, and the like.
Parenteral formulations may also be prepared in a dehydrated form (e.g., by lyophilization) or as sterile non-aqueous solutions. These formulations can be used with a suitable vehicle, such as sterile water. Solubility-enhancing agents may also be used in preparation of parenteral solutions.
Topical Administration
Compounds of the present invention may be administered topically to the skin or transdermally. Formulations for this topical administration can include lotions, solutions, creams, gels, hydrogels, ointments, foams, implants, patches and the like. Pharmaceutically acceptable carriers for topical administration formulations can include water, alcohol, mineral oil, glycerin, polyethylene glycol and the like. Topical administration can also be performed by electroporation, iontophoresis, phonophoresis and the like.
Compositions for topical administration may be formulated as immediate or modified release, including delayed or sustained release.
The present invention relates to compounds capable of exhibiting various pharmacological activities by inhibiting or degrading the activity of EGFR mutant proteins, pharmaceutical compositions comprising them as an active ingredient, and their medical uses (especially for the treatment or prevention of cancers showing EGFR mutations, especially lung cancer) and a treatment method comprising administering them to a subject in need of treatment or prevention. The compounds according to the present invention or pharmaceutically acceptable salts thereof are excellent in various aspects such as safety due to high selectivity for mutant EGFR, and may exhibit excellent medical effects in terms of activity inhibition or degradation of EGFR mutant.
Hereinafter, the present invention is described in considerable detail with examples to help those skilled in the art understand the present invention. However, the following examples are offered by way of illustration and are not intended to limit the scope of the invention. It is apparent that various changes may be made without departing from the spirit and scope of the invention or sacrificing all of its material advantages.
Preparation of Compounds of the Present Invention
Hereinafter, the synthesis process of some compounds of the present invention will be described, and the other compounds not mentioned below can be prepared by substituting starting materials, intermediates and/or reactants in a similar manner.
To a solution of 5-bromopentanoic acid (398 mg, 2.20 mmol) in dichloromethane (5 mL) was added oxalyl chloride (964 mL, 11.00 mmol), followed by 1 drop of N,N-dimethylformamide. The solution was concentrated for 1 hour. Tetrahydrofuran (10 mL) and 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (200 mg, 0.73 mmol) were added, and stirred overnight at 75° C. It was extracted by diluting with ethyl acetate and brine, and dried over anhydrous magnesium sulfate. After filtering and concentrating, the product was purified with a silica gel column to obtain the title compound (244.1 mg, 77%).
A mixture of 4-fluoro-2-methoxy-1-nitrobenzene (5.0 g, 29.22 mmol), Boc-piperazine (6.5 g, 35.06 mmol) and potassium carbonate (6.0 g, 43.24 mmol) in N,N-dimethylformamide (43 mL) was stirred at room temperature. After stirring overnight, potassium carbonate (3.0 g) was added and stirred for 2 days. Water was added to the reaction mixture, and the mixture was stirred, filtered, washed with water, and vacuum dried to obtain the title compound (9.5 g, 96%).
A mixture of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate (500 mg, 1.48 mmol) in tetrahydrofuran/ethanol (5/5 mL) was purged with argon gas and 10% Pd/C (50 mg) was added. After substituted with hydrogen gas, it was stirred for 4 hours, filtered, and concentrated to obtain the title compound (450 mg, 99%).
A mixture of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate (450 mg, 1.46 mmol), N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (508 mg, 1.46 mmol), BINAP (255 mg, 0.41 mmol) and potassium carbonate (464 mg, 3.37 mmol) in 1,4-dioxane (5 mL) was purged with argon gas and Pd(OAc)2 (50 mg, 0.20 mmol) was added. The mixture was stirred under reflux overnight, cooled to room temperature, and filtered through Celite. It was extracted by diluting with ethyl acetate and brine, and dried over anhydrous magnesium sulfate. After filtering and concentrating, the title compound (700 mg, 77%) was obtained by purification using a silica gel column.
Tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate (2.9 g, 4.68 mmol) was added to a reactor containing 20% trifluoroacetic acid/dichloromethane (46 mL), stirred for 3 hours, and neutralized with an aqueous solution of sodium bicarbonate. After extraction with dichloromethane, drying over anhydrous magnesium sulfate, filtration, and concentration, the title compound (2.4 g, 98%) was obtained.
To a solution of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide (67 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.15 mmol) in N,N-dimethylformamide (1.5 mL), diisopropylethylamine (67 mL, 0.39 mmol) was added and stirred at 90° C. After 16 hours, it was concentrated and purified by column to obtain the title compound (95 mg, 66%).
A mixture of 4-fluoro-2-methoxy-1-nitrobenzene (500 mg, 2.92 mmol), piperidone hydrochloride (470 mg, 3.47 mmol) and potassium carbonate (799 mg, 5.78 mmol) in dimethylsulfoxide (6 mL) was stirred overnight at 80° C. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed sequentially with water and saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated. The residue was purified by column to obtain the title compound (370 mg, 50%).
To a mixture of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one (460 mg, 1.84 mmol) and N-Boc-piperazine (1.0 g, 5.51 mmol) in dichloromethane (18 mL), acetic acid (0.3 mL, 5.51 mmol) was added and stirred for 1 hour and 30 minutes. After adding NaBH(OAc)3 (580 mg, 2.76 mmol) and stirring for 5 hours, the pH was adjusted to ˜8 with an aqueous solution of sodium hydrogen carbonate, followed by extraction with dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated, and the residue was purified by column to obtain the title compound (650 mg, 84%).
A mixture of tertiary butyl 4-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)piperazin-1-carboxylate (650 mg, 1.55 mmol) in tetrahydrofuran/ethanol (7/7 mL) was purged with argon gas, and 10% Pd/C (65 mg) was added. After replacing with hydrogen gas and stirring for 4 hours, the mixture was filtered through celite and concentrated to obtain the title compound (650 mg, 100%).
A mixture of tertiary butyl 4-(1-(4-amino-3-methoxyphenyl)piperidin-4-yl)piperazin-1-carboxylate (640 mg, 1.64 mmol), N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (570 mg, 1.64 mmol), BINAP (285 mg, 0.46 mmol) and potassium carbonate (521 mg, 3.77 mmol) in 1,4-dioxane (6.5 mL) was purged with argon gas, and Pd(OAc)2 (52 mg, 0.23 mmol) was added. After stirring under reflux overnight and cooling to room temperature, extraction was performed by diluting with ethyl acetate and brine. After drying over anhydrous magnesium sulfate, filtering and concentration, the title compound (610 mg, 53%) was obtained by purification with a silica gel column.
20% trifluoroacetic acid/dichloromethane (9 mL) was added to a reactor containing tertiary butyl 4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-carboxylate (610 g, 0.87 mmol), stirred for 3 hours, and then neutralized with an aqueous solution of sodium bicarbonate. After extraction with dichloromethane, drying over anhydrous magnesium sulfate, filtration, and concentration, the title compound (430 mg, 82%) was obtained.
To a solution of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide (51 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) in N,N-dimethylformamide (1.5 mL), diisopropylethylamine (51 mL, 0.29 mmol) was added and stirred at 90° C. After 16 hours, it was concentrated and purified by column to obtain the title compound (95 mg, 66%).
2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindolin-1,3-dione (500 mg, 1.82 mmol) was dissolved in tetrahydrofuran (9 mL) and 4-bromobutan-1-ol (0.289 mL, 2.74 mmol) and triphenylphosphine (717 mg, 2.74 mmol) were added. Diisopropyl azodicarboxylate (0.538 m, 2.74 mmol) was added slowly and stirred overnight. The reaction mixture was concentrated, diluted with isopropanol and stirred for 2 hours. The precipitated solid was filtered, washed with isopropanol, and dried to obtain the title compound (510 mg, 69%).
The title compound (3.90 g, 91%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with 4-fluoro-2-methoxy-1-nitrobenzene (2.0 g, 11.69 mmol) and tertiary butyl methyl(piperidin-4-yl)carbamate (2.76 g, 12.86 mmol) instead of 4-fluoro-2-methoxy-1-nitrobenzene and Boc-piperazine, respectively.
The title compound (1.65 g, yield 94%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)(methyl)carbamate (2.0 g, 7.99 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (1.52 g, yield 53%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl(1-(4-amino-3-methoxyphenyl)piperidin-4-yl)(methyl)carbamate (1.5 g, 4.47 mmol) and N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (1.63 g, 4.47 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate and N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (410 mg, yield 97%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)(methyl)carbamate (0.5 g, 0.774 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (21 mg, yield 16%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 4-(4-bromobutoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (60 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(methylamino)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (236 mg, yield 62%) was obtained using a method similar to the Step 1 synthesis method of Example 1 above with 11-bromoundecanoic acid (583 mg, 2.20 mmol) instead of 5-bromopentanoic acid.
The title compound (94 mg, yield 64%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 11-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)undecanamide (80 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (65 mg, yield 54%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 11-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)undecanamide (60 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
To a mixture of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (200 mg, 0.43 mmol), 5-bromopentanoic acid (155 mg, 0.86 mmol) and dimethylaminopyridine (11 mg, 0.086 mmol) in dichloromethane (4 mL), diisopropylethylamine (149 mL, 0.86 mmol) and stirred for 5 minutes, and then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (107 mg, 0.56 mmol) was added. After stirring overnight, it was diluted with water and extracted with dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated, and then purified with a silica gel column to obtain the title compound (203 mg, 80%).
The title compound (46 mg, yield 34%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with (2R, 4R)-1-((S)-2-(5-bromopentanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide (80 mg, 0.14 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.14 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (71 mg, yield 56%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 5-bromopentanoate (43 mg, 0.18 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.17 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The compound obtained in Step 1 was dissolved in 20% trifluoroacetic acid/dichloromethane (1 mL) and stirred overnight. The reaction mixture was concentrated, diluted with 4N 1,4-dioxane (1.5 mL), stirred for 1 hour, and then concentrated and used in the next reaction.
The title compound (60 mg, yield 57%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-(4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)pentanoic acid obtained in Step 2 instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentaoic acid, respectively.
The title compound (404 mg, yield 94%) was obtained using a method similar to the Step 1 synthesis method of Example 1 above with 2-(2-(2-chloroethoxy)ethoxy)acetic acid (535 mg, 2.93 mmol) and 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (268 mg, 0.98 mmol) instead of 5-bromopentanoic acid and 4-amino-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (200 mg, 0.73 mmol), respectively.
A mixture of 2-(2-(2-chloroethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide (400 mg, 0.91 mmol) and sodium iodide (1.3 g, 9.14 mmol) in acetone (6 mL) was stirred under reflux overnight. The reaction mixture was concentrated and extracted by diluting with water and dichloromethane. The organic layer was dried over anhydrous magnesium sulfate and filtered through silica gel. After concentration, the title compound (456 mg, 95%) was obtained by recrystallization from dichloromethane/methyl tertiary butyl ether/heptane.
The title compound (96 mg, yield 77%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-2-(2-(2-iodoethoxy)ethoxy)acetamide (71 mg, 0.14 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.14 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (59 mg, yield 51%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)-2-(2-(2-iodoethoxy)ethoxy)acetamide (61 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
A mixture of tertiary butyl 2-(2-(2-chloroethoxy)ethoxy)acetate (300 mg, 1.26 mmol) and sodium iodide (754 mg, 5.02 mmol) in acetone (8 mL) was stirred under reflux overnight. The reaction mixture was concentrated and extracted by diluting with water and dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by a column to obtain the title compound (387 mg, 93%).
The title compound (198 mg, yield 91%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 2-(2-(2-iodoethoxy)ethoxy)acetate (100 mg, 0.30 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (160 mg, 0.30 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 7 above with tertiary butyl 2-(2-(2-(4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)ethoxy)ethoxy)acetate (195 mg, 0.27 mmol) instead of tertiary butyl-5-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pentanoate, and then was used in the next reaction.
The title compound (99 mg, yield 41%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (105 mg, 0.23 mmol) and 2-(2-(2-(4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)ethoxy)ethoxy)acetic acid hydrochloride instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
To a solution of tertiary butyl 2-bromoacetate (34 mg, 0.17 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.17 mmol) in dichloromethane (1.5 mL), diisopropylethylamine (72 μL, 0.43 mmol) was added and stirred for 4 hours. Extraction was performed by dilution with water and dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated, and used in the next reaction.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 7 above with tertiary butyl 2-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)acetate (122 mg, 0.17 mmol) instead of tertiary butyl-5-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pentanoate, and then used in the next reaction.
The title compound (40 mg, yield 28%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (62 mg, 0.13 mmol) and 2-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)acetic acid hydrochloride instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (257 mg, yield 80%) was obtained using a method similar to the Step 1 synthesis method of Example 3 above with 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindolin-1,3-dione (200 mg, 0.73 mmol) and 2-(2-chloroethoxy)ethan-1-ol (136 mg, 1.10 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindolin-1,3-dione and 4-bromobutan-1-ol, respectively.
The title compound (215 mg, yield 71%) was obtained using a method similar to the Step 2 synthesis method of Example 8 above with 4-(2-(2-chloroethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (250 mg, 0.57 mmol) instead of 2-(2-(2-chloroethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide.
The title compound (65 mg, yield 50%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2,6-dioxopiperidin-3-yl)-4-(2-(2-iodoethoxy)ethoxy)isoindolin-1,3-dione (80 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (79 mg, yield 72%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2,6-dioxopiperidin-3-yl)-4-(2-(2-iodoethoxy)ethoxy)isoindolin-1,3-dione (61 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (25 mg, yield 20%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 4-(4-bromobutoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (60 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (76 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (23 mg, yield 17%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 4-(4-bromobutoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (60 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (88 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
To a solution of 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (500 mg, 1.81 mmol) in dimethylformamide (1.5 mL), 2-(2-aminoethoxy)ethane-1-ol (209 mg, 1.99 mmol) and N,N-diisopropylethylamine (64 μl, 0.368 mmol) were added and stirred at 90° C. overnight. Extraction was performed by diluting with ethyl acetate and water, and the organic layer was washed with saturated brine. After drying with anhydrous magnesium sulfate, filtration, and concentration, the title compound (251 mg, 39%) was obtained by column purification.
2-(2,6-dioxopiperidin-3-yl)-4-((2-(2-hydroxyethoxy)ethyl)amino)isoindolin-1,3-dione (150 mg, 0.42 mmol) and 4-toluenesulfonyl chloride (95 mg, 0.50 mmol) were dissolved in dichloromethane (4 mL) and triethylamine (0.17 mL, 1.25 mmol) was slowly added at 0° C. After stirring at room temperature for 24 hours, the mixture was washed with water, and the organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated. The residue was purified by column to obtain the title compound (155 mg, yield 73%).
The title compound (29 mg, yield 25%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethoxy)ethyl 4-methylbenzenesulfonate (70 mg, 0.14 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.14 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title Compound (2.0 g, yield 98%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with 1,2-difluoro-4-nitrobenzene (1.0 g, 6.29 mmol) instead of 4-fluoro-2-methoxy-1-nitrobenzene.
The title compound (430 mg, yield 95%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 4-(2-fluoro-4-nitrophenyl)piperazin-1-carboxylate (500 mg, 1.54 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (540 mg, yield 56%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 4-(4-amino-2-fluorophenyl)piperazin-1-carboxylate (450 mg, 1.52 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (70 mg, yield 84%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperazin-1-carboxylate (100 mg, 0.17 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (41 mg, yield 34%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with N-(2-((5-chloro-2-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (59 mg, 0.12 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (600 mg, yield 80%) was obtained using a method similar to the Step 1 synthesis method of Example 2 above with 1,2-difluoro-4-nitrobenzene (500 mg, 3.14 mmol) instead of 4-fluoro-2-methoxy-1-nitrobenzene.
The title compound (320 mg, yield 62%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with 1-(2-fluoro-4-nitrophenyl)piperidin-4-one (300 mg, 1.26 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one.
The title compound (300 mg, yield 100%) was obtained using a method similar to the Step 3 synthesis method of Example 2 above with tertiary butyl 4-(1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)piperazin-1-carboxylate (320 mg, 0.78 mmol) instead of tertiary butyl 4-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)piperazin-1-carboxylate.
The title compound (280 mg, yield 53%) was obtained using a method similar to the Step 3 synthesis method of Example 2 above with tertiary butyl 4-(1-(4-amino-2-fluorophenyl)piperidin-4-yl)piperazin-1-carboxylate (290 mg, 0.77 mmol) instead of tertiary butyl 4-(1-(4-amino-3-methoxyphenyl)piperidin-4-yl)piperazin-1-carboxylate.
The title compound (2.5 g, yield 96%) was obtained using a method similar to the Step 3 synthesis method of Example 2 above with tertiary butyl 4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperidin-4-yl)piperazin-1-carboxylate (3.1 g, 4.43 mmol) instead of tertiary butyl 4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-carboxylate.
The title compound (42 mg, yield 32%) was obtained using a method similar to the Step 2 synthesis method of Example 6 above with N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound was obtained using a method similar to the Step 1 synthesis method of Example 11 above with N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.17 mmol) instead of tertiary butyl 2-bromoacetate (34 mg, 0.17 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, and was used in the next reaction.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 7 above with tertiary butyl 2-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperidin-4-yl)piperazin-1-yl)acetate (119 mg, 0.17 mmol) instead of tertiary butyl-5-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)pentanoate, and then was used in the next reaction.
The title compound (80 mg, yield 36%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (62 mg, 0.13 mmol) and 2-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperidin-4-yl)piperazin-1-yl)acetic acid hydrochloride instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
To a solution of tertiary butyl 3-bromopropanoate (35 mg, 0.17 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.17 mmol) in dichloromethane (1.5 mL), diisopropylethylamine (74 mL, 0.43 mmol) was added and stirred for 4 hours. Extraction was performed by dilution with water and dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated, and used in the next reaction.
To tertiary butyl 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoate obtained in Example 20-1 Step above, 4N HCl 1,4-dioxane solution (2 mL) and dichloromethane (2 mL) were added and stirred at 40° C. for 3 hours. The title compound obtained by concentrating and vacuum drying the reaction mixture was used in the next reaction.
The title compound (57 mg, yield 37%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (75 mg, 0.16 mmol) and 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (66 mg, yield 63%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide (46 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (72 mg, yield 69%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)acetamide (44 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (647 mg, yield 42%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 4-(piperidin-4-ylmethyl)piperazin-1-carboxylate (1.0 g, 3.54 mmol) instead of Boc-piperazine.
The title compound (572 mg, yield 96%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 4-((1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (640 mg, 1.47 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (625 mg, yield 63%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 4-((1-(4-amino-2-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (570 mg, 1.41 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (535 mg, yield 100%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 4-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (620 mg, 0.17 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (70 mg, yield 67%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide (45 mg, 0.11 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.11 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (78 mg, yield 75%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)acetamide (43 mg, 0.11 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.11 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (903 mg, yield 61%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with benzyl 4-(piperazin-1-ylmethyl)piperidin-1-carboxylate (1.0 g, 3.16 mmol) instead of Boc-piperazine.
To a solution of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (900 mg, 1.92 mmol) in 1,4-dioxane (15 mL), NHCl (513 mg, 9.60 mmol) in H2O (5 mL) was added and purged with argon. Zn (628 mg, 9.60 mmol) was added and stirred at 70° C. After 19 hours, the mixture was filtered through Celite and concentrated. The residue was diluted with aqueous sodium chloride solution, made basic with aqueous saturated sodium hydrocarbon solution, and extracted with ethyl acetate. The organic layer was dried over sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound (831 mg, yield 99%).
The title compound (1.0 g, yield 71%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with benzyl 4-((4-(4-amino-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (830 mg, 1.89 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (617 mg, yield 94%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with benzyl 4-((4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (800 mg, 1.06 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (33 mg, yield 31%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide (45 mg, 0.11 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.11 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (78 mg, yield 75%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)acetamide (43 mg, 0.11 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.11 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (119 mg, yield 100%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 2-bromoacetate (32 mg, 0.16 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.16 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl-2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetate (119 mg, 0.16 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
The title compound (104 mg, yield 59%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid hydrochloride obtained in Step 2 instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (91 mg, yield 94%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 2-bromoacetate (26 mg, 0.13 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (82 mg, 0.13 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl-2-(4-((4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (91 mg, 0.13 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
The title compound (73 mg, yield 51%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 2-(4-((4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)acetic acid hydrochloride obtained in Step 2 instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (462 mg, yield 64%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 1-(piperidin-4-ylmethyl)piperidin-4-carboxylate (470 mg, 1.66 mmol) instead of Boc-piperazine.
The title compound (223 mg, yield 99%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 1-((1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (240 mg, 0.55 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (95 mg, yield 24%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 1-((1-(4-amino-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (223 mg, 0.55 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (90 mg, 0.13 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate, and then used in the next reaction.
The title compound (27 mg, yield 20%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride obtained in Step 2 instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (13.2 g, yield 93%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 3,9-diazaspiro[5,5]undecan-3-carboxylate (6.0 g, 35.06 mmol) instead of Boc-piperazine.
The title compound (1.4 g, yield 99%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 9-(3-methoxy-4-nitrophenyl)-3,9-diazaspiro[5,5]undecan-3-carboxylate (1.5 g, 3.70 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (1.4 g, yield 54%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 9-(4-amino-3-methoxyphenyl)-3,9-diazaspiro[5,5]undecan-3-carboxylate (1.4 g, 3.60 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (1.2 g, yield 99%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 9-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)-3,9-diazaspiro[5,5]undecan-3-carboxylate (1.4 g, 1.98 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (61 mg, yield 56%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 3-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)propanamide (49 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(3,9-diazaspiro[5,5]undecan-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (70 mg, 0.12 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (119 mg, yield 100%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 2-bromoacetate (26 mg, 0.13 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(3,9-diazaspiro[5,5]undecan-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (100 mg, 0.17 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl-2-(4-((4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)acetate (119 mg, 0.17 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
The title compound (85 mg, yield 34%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 2-(9-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)-3,9-diazaspiro[5,5]undecan-3-yl)acetic acid hydrochloride obtained in Step 2 instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (4.8 g, yield 79%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 2,6-diazaspiro[3,3]undecan-2-carboxylate (7.4 g, 21.0 mmol) instead of Boc-piperazine.
The title compound (950 mg, yield 70%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 6-(3-methoxy-4-nitrophenyl)-2,6-diazaspiro[3,3]heptan-2-carboxylate (1.5 g, 4.29 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (0.7 g, yield 34%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 6-(4-amino-3-methoxyphenyl)-2,6-diazaspiro[3,3]heptan-2-carboxylate (0.9 g, 2.81 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (580 mg, yield 99%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 6-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)-2,6-diazaspiro[3,3]undecan-2-carboxylate (0.7 g, 1.98 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (48 mg, yield 41%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 3-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)propanamide (54 mg, 0.13 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(2,6-diazaspiro[3,3]heptan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (70 mg, 0.13 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
Sodium hydroxide (346 mg, 8.66 mmol) was added to a solution of tertiary butyl 3-hydroxyazetidin-1-carboxylate (1.0 g, 5.77 mmol) in DMF (11 mL) and stirred for 30 minutes. ((2-bromoethoxy)methyl)benzene (1.1 mL, 6.93 mmol) was added slowly at 0° C. After stirring overnight at 80° C., extraction was performed with ethyl acetate and water. The organic layer was washed sequentially with water and saturated brine, and dried over anhydrous magnesium sulfate. The title compound (0.95 g, yield 54%) was obtained by filtration, concentration and purification by column chromatography.
A solution of tertiary butyl 3-(2-(benzyloxy)ethoxy)azetidin-1-carboxylate (0.9 g, 2.93 mmol) in 20% trichloroacetic acid in dichloromethane (20 mL) was stirred at room temperature for 5 hours. The reaction mixture was concentrated and extracted with 1N aqueous sodium hydroxide solution and dichloromethane. The organic layer was dried over sodium sulfate, filtered, and concentrated to obtain the title compound (550 mg, yield 92%).
To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione (0.6 g, 2.17 mmol) and 3-(2-(benzyloxy)ethoxy)azetidine (0.5 g, 2.39 mmol) in N,N-dimethylformamide (10 mL), diisopropylethylamine (757 mL, 4.34 mmol) was added and stirred overnight at 80° C. After extraction with ethyl acetate and water, the organic layer was washed with saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain the title compound (650 mg, yield 65%).
A solution of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (500 mg, 1.08 mmol) in tetrahydrofuran/methanol (1/1, 20 mL) was purged with argon and 10% Pd/C (0.2 g) was added. The reaction mixture was substituted with hydrogen and stirred overnight. It was diluted with dichloromethane, filtered through celite, and concentrated. The residue was purified by column chromatography to obtain the title compound (130 mg, yield 32%).
A solution of 2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-hydroxyethoxy)azetidin-1-yl)isoindolin-1,3-dione (130 mg, 0.35 mmol) in dichloromethane (7 mL), triethylamine (146 mL, 1.05 mmol) was added and then methanesulfonyl chloride (54 mL, 0.70 mmol) was slowly added at 0° C. The reaction mixture was extracted with water and dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to obtain the title compound (90 mg, yield 57%).
The title compound (40 mg, yield 31%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)oxy)ethylmethane sulfonate (70 mg, 0.16 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(2,6-diazaspiro[3,3]heptan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (81 mg, 0.16 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (0.7 g, yield 42%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with tertiary butyl 4-hydroxypiperidin-1-carboxylate (1.0 g, 4.97 mmol) instead of tertiary butyl 3-hydroxyazetidin-1-carboxylate.
The title compound (480 mg, yield 98%) was obtained using a method similar to the Step 2 synthesis method of Example 33 above with tertiary butyl 4-(2-(benzyloxy)ethoxy)piperidin-1-carboxylate (0.7 g, 2.09 mmol) instead of tertiary butyl 3-(2-(benzyloxy)ethoxy)azetidin-1-carboxylate.
The title compound (400 mg, yield 45%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 4-(2-(benzyloxy)ethoxy)piperidine (468 mg, 1.99 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (130 mg, yield 40%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with 5-(4-(2-(benzyloxy)ethoxy)piperidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (0.4 g, 0.81 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (135 mg, yield 90%) was obtained using a method similar to the Step 5 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-hydroxyethoxy)piperidin-1-yl)isoindolin-1,3-dione (125 mg, 0.31 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-hydroxyethoxy)azetidin-1-yl)isoindolin-1,3-dione.
The title compound (70 mg, yield 53%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)oxy)ethylmethanesulfonate (70 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(2,6-diazaspiro[3,3]heptan-2-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (76 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (428 mg, yield 72%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with 2-chloro-4-fluoro-1-nitrobenzene (221 mg, 1.26 mmol) instead of 4-fluoro-2-methoxy-1-nitrobenzene.
The title compound (420 mg, yield 99%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with 4-((4-(3-chloro-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (408 mg, 0.91 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
To a solution of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (420 mg, 0.95 mmol) and N-(2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (396 mg, 1.14 mmol) in isopropanol (9.5 mL), methanesulfonic acid (0.12 mL, 1.90 mmol) was added and stirred under reflux overnight. The mixture was extracted by diluting with dichloromethane and aqueous sodium hydroxide solution, and dried over anhydrous sodium sulfate. After filtering and concentrating, the title compound (316 mg, 44%) was obtained by purification using a silica gel column.
A saturated aqueous solution of benzyl 4-((4-(3-chloro-4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (50 mg, 0.066 mmol) in hydrochloric acid (1 mL) was stirred overnight at room temperature. The mixture was extracted by diluting with dichloromethane and aqueous sodium hydroxide solution, and dried over anhydrous sodium sulfate. The title compound (21 mg, 52%) was obtained by filtration and concentration.
The title compound (50 mg, yield 99%) was obtained using a method similar to the Step 1 synthesis method of Example 11 above with N-(2-((5-chloro-2-((2-chloro-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (71 mg, 0.114 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 2-(4-((1-(3-chloro-4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetate (50 mg, 0.068 mmol) instead of tertiary butyl 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoate, and then used in the next reaction.
The title compound (51 mg, yield 69%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-(4-((1-(3-chloro-4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)phenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid hydrochloride (46 mg, 0.068 mmol) instead of 5-bromopentanoic acid.
The title compound (765 mg, yield 67%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with 1,2-difluoro-4-nitrobenzene (400 mg, 2.51 mmol) and benzyl 4-(piperazin-1-ylmethyl)piperidin-1-carboxylate (798 mg, 2.51 mmol) instead of 4-fluoro-2-methoxy-1-nitrobenzene and Boc-piperazine, respectively.
The title compound (700 mg, yield 99%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with benzyl 4-((4-(2-fluoro-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (750 mg, 1.64 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (1.1 g, 91%) was obtained using a method similar to the Step 3 synthesis method of Example 35 above with benzyl 4-((4-(4-amino-2-fluorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (700 mg, 1.64 mmol) instead of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (900 mg, yield 99%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (1.1 g, 1.49 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (75 mg, yield 63%) was obtained using a method similar to the Step 1 synthesis method of Example 11 above with N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.166 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (65 mg, 98%) was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetate (70 mg, 0.098 mmol) instead of tertiary butyl 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoate.
The title compound (32 mg, yield 31%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-2-fluorophenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid hydrochloride (45 mg, 0.096 mmol) instead of 5-bromopentanoic acid.
The title compound (230 mg, yield 87%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 3-formylazetidin-1-carboxylate (107 mg, 0.58 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (200 mg, 0.39 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 3-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)azetidin-1-carboxylate (225 mg, 0.33 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (40 mg, yield 42%) was obtained using a method similar to the Step 1 synthesis method of Example 11 above with N-(2-((2-((4-(4-(azetidin-3-ylmethyl)piperazin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.166 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 2-(3-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)azetidin-1-yl)acetate (80 mg, 0.117 mmol) instead of tertiary butyl 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoate.
The title compound (45 mg, yield 48%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-(3-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)azetidin-1-yl)acetic acid hydrochloride (85 mg, 0.118 mmol) instead of 5-bromopentanoic acid.
The title compound (640 mg, yield 56%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with benzyl 4-formylpiperidin-1-carboxylate (732 mg, 2.96 mmol) and tertiary butyl azetidin-3-carboxylate (857 mg, 4.44 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (400 mg, yield 95%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((3-(tertiary butoxycarbonyl)azetidin-1-yl)methyl)piperidin-1-carboxylate (1.3 g, 3.40 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (510 mg, yield 80%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 1-(piperidin-4-ylmethyl)azetidin-3-carboxylate (400 mg, 1.57 mmol) instead of Boc-piperazine.
The title compound (470 mg, yield 99%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with benzyl tertiary butyl 1-((1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)methyl)azetidin-3-carboxylate (510 mg, 1.26 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (506 mg, yield 59%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 1-((1-(4-amino-3-methoxyphenyl)piperidin-4-yl)methyl)azetidin-3-carboxylate (470 mg, 2.25 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)azetidin-3-carboxylate (100 mg, 0.146 mmol) instead of tertiary butyl 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoate.
The title compound (87 mg, yield 60%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 1-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)azetidin-3-carboxylic acid hydrochloride (87 mg, 0.14 mmol) instead of 5-bromopentanoic acid.
A mixture of 3-(4-hydroxy-1-oxoisoindolin-2-yl)piperidin-2,6-dione (297 mg, 1.14 mmol), tertiary butyl 3-iodoazetidin-1-carboxylate (646 mg, 2.28 mmol) and potassium carbonate (315 mg, 2.28 mmol) in N,N-dimethylformamide (3 mL) was stirred at 70° C. After stirring overnight, tertiary butyl 3-iodoazetidin-1-carboxylate (200 mg) was further added and stirred for 4 hours. A saturated NH4Cl aqueous solution was added to the reaction mixture, stirred, filtered, washed with water and ethyl acetate, and vacuum dried to obtain the title compound (150 mg, 36%).
The title compound was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)oxy)azetidin-1-carboxylate (150 mg, 0.36 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate, and then used in the next reaction.
To a solution of 3-(4-(azetidin-3-yloxy)-1-oxoisoindolin-2-yl)piperidin-2,6-dione hydrochloride (127 mg, 0.36 mmol) and 1-bromo-2-chloroethane (33 μL, 0.40 mmol) in N,N-dimethylformamide (3.5 mL), diisopropylethylamine (219 μL, 0.29 mmol) was added and stirred at 90° C. After 4.5 hours, it was concentrated and purified by column to obtain the title compound (70 mg, 51%).
A mixture of 3-(4-((1-(2-chloroethyl)azetidin-3-yl)oxy)-1-oxoisoindolin-2-yl)piperidin-2,6-dione (70 mg, 0.185 mmol), N-(2-((5-chloro-2-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (93 mg, 0.185 mmol, sodium iodide (14 mg, 0.093 mmol), sodium hydroxide (31 mg, 0.371 mmol) and tetrabutylammonium bromide (TBAB, 30 mg, 0.093 mmol) in N,N-dimethylformamide (1 mL) was stirred overnight at 80° C. The mixture was cooled to room temperature, water was added, and a solid was obtained by filtration, and then the title compound (35 mg, 23%) was obtained by column purification.
The title compound (57 mg, yield 43%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5yl)azetidin-3-yl)oxy)ethylmethanesulfonate (70 mg, 0.16 mmol) and N-(2-((5-chloro-2-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (78 mg, 0.16 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
A mixture of 5-methylisofuran-1,3-dione (5.0 g, 30.84 mmol), 3-aminopiperidin-2,6-dione (6.1 g, 37.01 mmol) and sodium acetate (2.5 g, 30.84 mmol) in acetic acid (50 mL) was stirred at 80° C. After 6 hours, the mixture was cooled to room temperature, water was added, and the mixture was stirred. The precipitated solid was filtered, washed with methyl tertiary butyl ether and dried to obtain the title compound (6.1 g, 72%).
To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-methylisoindolin-1,3-dione (200 mg, 0.74 mmol) and NBS (144 mg, 0.81 mmol) in chloroform (10 mL), AIBN (24 mg, 0.15 mmol) was added and stirred under reflux. After stirring overnight, the mixture was extracted with water, dried over anhydrous magnesium sulfate, and filtered, and the obtained material (140 mg, mixture) was used in the next reaction.
To a solution of 5-(bromomethyl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (44 mg, 0.125 mmol) and N-(2-((5-chloro-2-((2 N,N of -methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.114 mmol) in N,N-dimethylformamide (1 mL), DIPEA (50 μL, 0.285 mmol) was added and stirred at 50° C. overnight. After cooling to room temperature, water was added and filtered to obtain a solid, which was purified by column to obtain the title compound (48 mg, 48%).
The title compound (56 mg, yield 55%) was obtained using a method similar to the Step 3 synthesis method of Example 41 above with N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.119 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (30 mg, yield 28%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (50 mg, 0.151 mmol) and N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.116 mmol) instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (120 mg, yield 93%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with (S)-pyrrolidin-2-ylmethanol (44 mg, 0.43 mmol) instead of 3-(2-(benzyloxy)ethoxy)axetidine.
The title compound (416 mg, yield 38%) was obtained using a method similar to the Step 5 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-5-((S)-2-(hydroxymethyl)pyrrolidin-1-yl)isoindolin-1,3-dione (1.0 g, 2.53 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-hydroxyethoxy)azetidin-1-yl)isoindolin-1,3-dione.
The title compound (18 mg, yield 17%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with ((2S)-1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoindolin-5-yl)pyrrolidin-2-yl)methyl methanesulfonate (50 mg, 0.115 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (69 mg, 0.115 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
A mixture of 2-(2-bromoethyl)isoindolin-1,3-dione (205 mg, 0.81 mmol), N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (500 mg, 0.81 mmol) and potassium carbonate (280 mg, 2.03) in acetonitrile (8 mL) was stirred overnight at 70° C. The reaction mixture was concentrated and extracted with water and dichloromethane. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography to obtain the title compound (350 mg, yield 57%).
To a suspension of N-(2-((5-chloro-2-((4-(4-(4-(2-(1,3-dioxoindolin-2-yl)ethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (350 mg, 0.45 mmol) in ethanol (4.5 mL), hydrazine hydrate (57 mg, 1.13 mmol) was added and stirred at 80° C. for 4 hours. The reaction mixture was concentrated and extracted with water and dichloromethane. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to quantitatively obtain the title compound, which was used in the next reaction.
The title compound (35 mg, yield 36%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (30 mg, 0.109 mmol) and N-(2-((2-((4-(4-(4-(2-aminoethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (71 mg, 0.109 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (323 mg, yield 94%) was obtained using a method similar to the Step 1 synthesis method of Example 3 above with 2-chloroethan-1-ol (110 μL, 1.64 mmol) instead of 4-bromobutan-1-ol.
The title compound (341 mg, yield 78%) was obtained using a method similar to the Step 2 synthesis method of Example 8 above with 4-(2-chloroethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (320 mg, 1.01 mmol) instead of 2-(2-(2-chloroethoxy)ethoxy)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide.
The title compound (36 mg, yield 35%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2,6-dioxopiperidin-3-yl)-4-(2-iodoethoxy)isoindolin-1,3-dione (54 mg, 0.128 mmol) and N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.116 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (63 mg, yield 61%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)acetamide (44 mg, 0.116 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.116 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (63 mg, yield 60%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)acetamide (44 mg, 0.116 mmol) and N-(2-((5-chloro-2-((3-fluoro-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.116 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (700 mg, yield 43%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with benzyl 4-formylpiperidin-1-carboxylate (1.0 g, 4.04 mmol) and tertiary butyl piperidin-4-carboxylate (1.1 g, 6.07 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (470 mg, yield 99%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((4-(tertiary butoxycarbonyl)piperidin-1-yl)methyl)piperidin-1-carboxylate (700 mg, 1.68 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (240 mg, yield 12%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with tertiary butyl 1-(piperidin-4-ylmethyl)piperidin-4-carboxylate (1.3 g, 4.60 mmol) instead of Boc-piperazine.
The title compound (223 mg, yield 99%) was obtained using a method similar to the Step 3 synthesis method of Example 1 above with tertiary butyl 1-((1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (240 mg, 0.55 mmol) instead of tertiary butyl 4-(3-methoxy-4-nitrophenyl)piperazin-1-carboxylate.
The title compound (95 mg, yield 24%) was obtained using a method similar to the Step 4 synthesis method of Example 1 above with tertiary butyl 1-((1-(4-amino-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (223 mg, 0.55 mmol) instead of tertiary butyl 4-(4-amino-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylate (100 mg, 0.14 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
To a mixture of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride (97 mg, 0.14 mmol), 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione (36 mg, 0.86 mmol) and HATU (107 mg, 0.28 mmol) in N,N-dimethylformamide (1 mL), diisopropylethylamine (98 μL, 0.56 mmol) was added and stirred for 2 days. The reaction mixture was diluted with water and extracted with dichloromethane. The organic layer was dried over anhydrous magnesium sulfate, filtered, concentrated, and then purified with a silica gel column to obtain the title compound (21 mg, 17%).
The title compound (90 mg, yield 64%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)acetamide (60 mg, 0.158 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(3,9-diazaspiro[5.5]undecan-3-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (93 mg, 0.158 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (0.99 g, yield 89%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(piperidin-4-yl)ethan-1-ol (412 mg, 3.19 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (510 mg, yield 43%) was obtained using a method similar to the Step 2 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-hydroxyethyl)piperidin-1-yl)isoindolin-1,3-dione (0.99 g, 2.57 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-hydroxyethoxy)azetidin-1-yl)isoindolin-1,3-dione.
The title compound (30 mg, yield 29%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)ethyl methanesulfonate (50 mg, 0.108 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (65 mg, 0.108 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (54 mg, yield 52%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2,6-dioxopiperidin-3-yl)-4-(2-iodoethoxy)isoindolin-1,3-dione (50 mg, 0.116 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.116 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (42 mg, yield 41%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (42 mg, 0.125 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (70 mg, 0.114 mmol) instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
A mixture of tri-tertiary butylphosphonium tetrafluoroborate (TTBP·HBF4, 110 mg, 0.38 mmol), Pd2(dba)3 (163 mg, 0.18 mmol) and N-cyclohexyl-N-cyclohexanamine (0.27 mL, 1.28 mmol) in 1,4-dioxane (6 mL) was purged with argon gas and stirred for 30 minutes. (4-bromo-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (0.3 g, 0.89 mmol) and benzyl acrylate (0.3 mL, 1.98 mmol) were added and stirred at 55° C. for 12 hours. The reaction mixture was filtered through celite, concentrated, and purified by column chromatography to obtain the title compound (368 mg, yield 99%).
To a solution of benzyl (E)-3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acrylate (137 mg, 0.33 mmol) in tetrahydrofuran (5 mL), 10% Pd/C (27 mg) was added and and replaced with hydrogen gas. After stirring overnight, the mixture was filtered through celite and concentrated to obtain the title compound (105 mg, yield 96%).
The title compound (105 mg, yield 75%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)propanoic acid (50 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (92 mg, 0.15 mmol) instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid, respectively.
The title compound (330 mg, 57%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with (S)-2-((tertiary butoxycarbonyl)amino)-2-cyclopropylacetic acid (243 mg, 1.13 mmol) and (2R,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide (400 mg, 1.13 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl ((S)-1-cyclopropyl-2-((2R,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-2-oxoethyl)carbamate (324 mg, 0.63 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
The title compound (73 mg, 51%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with (2R,4R)-1-((S)-2-amino-2-cyclopropylacetyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboximide hydrochloride (60 mg, 0.134 mmol) and 2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid hydrochloride (100 mg, 0.134 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (232 mg, 79%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with (tertiary butoxycarbonyl)-L-valine (122 mg, 0.565 mmol) and (2R,4R)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide (200 mg, 0.565 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound was obtained using a method similar to the Step 2 synthesis method of Example 20 above with tertiary butyl ((S)-1-((2R,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3-methyl-1-oxobutan-2-yl)carbamate (460 mg, 0.89 mmol) instead of 3-(4-(1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)propanoic acid hydrochloride, and then used in the next reaction.
The title compound (63 mg, 44%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with (2R,4R)-1-(L-valyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride (61 mg, 0.134 mmol) and 2-(4-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperazin-1-yl)acetic acid hydrochloride (100 mg, 0.134 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (30 mg, yield 40%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(2,6-dioxopiperidin-3-yl)-4-(2-iodoethoxy)isoindolin-1,3-dione (34 mg, 0.079 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (50 mg, 0.079 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (20 mg, yield 20%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((2-((4-(4-(4-(2-aminoethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (71 mg, 0.109 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (340 mg, yield 97%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with benzyl 4-formylpiperidin-1-carboxylate (121 mg, 0.49 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (250 mg, 0.41 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (247 mg, yield 79%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((4-((4-(4-((5-chloro-4-((2-N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)methyl)piperidin-1-carboxylate (340 mg, 0.40 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (73 mg, yield 53%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-((1-(piperidin-4-ylmethyl)piperidin-4-yl)methyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.14 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (220 mg, yield >100%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 3-formylazetidin-1-carboxylate (54 mg, 0.293 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.244 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (250 mg, yield >100%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 3-((4-(1-(4-((5-chloro-4-((2-N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)azetidin-1-carboxylate (220 mg, 0.244 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (28 mg, yield 16%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((2-((4-(4-(4-(azetidin-3-ylmethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.189 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (220 mg, yield >100%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 3-formylazetidin-1-carboxylate (54 mg, 0.293 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.244 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine.
The title compound (260 mg, yield >100%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 3-((4-((4-(4-((5-chloro-4-((2-N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)methyl)azetidin-1-carboxylate (220 mg, 0.244 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (30 mg, yield 17%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((2-((4-(4-((1-(azetidin-3-ylmethyl)piperidin-4-yl)methyl)piperazin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.189 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (1.5 g, yield 80%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 4-formylpiperidin-1-carboxylate (945 mg, 4.43 mmol) and benzyl (piperidin-4-ylmethyl)carbamate (1.0 g, 4.03 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (900 mg, yield 81%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 4-((4-((((benzyloxy)carbonyl)amino)methyl)piperidin-1-yl)methyl)piperidin-1-carboxylate (1.5 g, 3.37 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (540 mg, yield 94%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with benzyl ((1-(piperidin-4-ylmethyl)piperidin-4-yl)methyl)carbamate (400 mg, 1.16 mmol) instead of Boc-piperazine.
The title compound (480 mg, yield 95%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with benzyl ((1-((1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)methyl)piperidin-4-yl)methyl)carbamate (540 mg, 1.09 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (480 mg, yield 60%) was obtained using a method similar to the Step 3 synthesis method of Example 35 above with benzyl ((1-((1-(4-amino-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-yl)methyl)carbamate (480 mg, 1.03 mmol) instead of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (420 mg, 0.95 mmol).
The title compound (395 mg, yield 99%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl ((1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-yl)methyl)carbamate (480 mg, 0.62 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (48 mg, yield 17%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (129 mg, 0.47 mmol) and N-(2-((2-((4-(4-((4-(aminomethyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (200 mg, 0.31 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine, respectively.
The title compound (65 mg, yield 47%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((2-((4-(4-((4-(aminomethyl)piperidin-1-yl)methyl)piperidin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.16 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (120 mg, yield 96%) was obtained using a method similar to the Step 1 synthesis method of Example 54 above with (5-bromo-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (100 mg, 0.30 mmol) instead of (4-bromo-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (60 mg, yield 63%) was obtained using a method similar to the Step 2 synthesis method of Example 54 above with benzyl (E)-3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)acrylate (120 mg, 0.29 mmol) instead of benzyl (E)-3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acrylate.
The title compound (75 mg, yield 67%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)propanoic acid (40 mg, 0.12 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (74 mg, 0.12 mmol) instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid.
The title compound (350 mg, 79%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (351 mg, 0.57 mmol) and (tertiary butoxycarbonyl)glycine (100 mg, 0.57 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (120 mg, yield 92%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl (2-(4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-oxoethyl)carbamate (150 mg, 0.19 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (80 mg, yield 53%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (45 mg, 0.16 mmol) and N-(2-((5-chloro-2-((4-(4-((1-glycylpiperidin-4-yl)methyl)piperazin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (110 mg, 0.16 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine, respectively.
The title compound (28 mg, yield 19%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((4-(4-((1-glycylpiperidin-4-yl)methyl)piperazin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (110 mg, 0.16 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (90 mg, yield 57%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (111 mg, 0.18 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (110 mg, yield 70%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (111 mg, 0.18 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (80 mg, 53%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (137 mg, 0.22 mmol) and (S)-hydroxypropanoic acid (20 mg, 0.22 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (60 mg, yield 55%) was obtained using a method similar to the Step 1 synthesis method of Example 3 above with (S)—N-(2-((5-chloro-2-((4-(4-((1-(2-hydroxypropanoyl)piperidin-4-yl)methyl)piperazin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.12 mmol) instead of 4-bromobutan-1-ol.
The title compound (130 mg, 79%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (130 mg, 0.21 mmol) and (tertiary butoxycarbonyl)-L-alanine (40 mg, 0.21 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (105 mg, yield 94%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl (S)-(1-(4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-1-oxopropan-2-yl))carbamate (128 mg, 0.16 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (60 mg, yield 60%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (30 mg, 0.11 mmol) and N-(2-((2-((4-(4-((1-(L-alanyl)piperidin-4-yl)methyl)piperazin-1-yl)-2-methoxyphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (75 mg, 0.11 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine, respectively.
The title compound (103 mg, yield 60%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 3-formylazetidin-1-carboxylate (65 mg, 0.31 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (130 mg, 0.22 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (90 mg, yield >100%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 4-((4-(1-(4-((5-chloro-4-((2-N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)piperazin-1-yl)methyl)piperidin-1-carboxylate (100 mg, 0.125 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (64 mg, yield 53%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-4-(4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (88 mg, 0.126 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
To a mixture of 5-bromo-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (3.0 g, 8.90 mmol), prop-2-yn-1-ol (0.78 mL, 12.82 mmol) and CuI (576 mg, 3.03 mmol) in N,N-dimethylformamide (45 mL), N,N-diisopropyldiamine (3.1 mL, 17.80 mmol) was added and purged with argon gas. Pd(PPh3)2Cl2 (1.0 g, 1.42 mmol) was added thereto and stirred overnight at 75° C. After extraction with dichloromethane and water, the organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated. The residue was recrystallized from toluene to obtain the title compound (1.0 g, yield 36%).
To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(3-hydroxyprop-1-yn-1-yl)isoindolin-1,3-dione (170 mg, 0.54 mmol) in N,N-dimethylformamide (5 mL), N,N-diisopropyldiamine (0.13 mL, 1.62 mmol) was added and then methanesulfonyl chloride (0.05 mL, 0.65 mmol) was slowly added at 0° C. After stirring the reaction mixture overnight, it was extracted with ethyl acetate and water. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by column chromatography to obtain the title compound (150 mg, yield 84%).
The title compound (180 mg, yield 87%) was obtained using a method similar to the Step 3 synthesis method of Example 41 above with 5-(3-chloroprop-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (77 mg, 0.23 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (140 mg, 0.23 mmol) instead of 5-(bromomethyl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (68 mg, yield 56%) was obtained using a method similar to the Step 3 synthesis method of Example 41 above with 5-(3-chloroprop-1-yn-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione (50 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (78 mg, 0.15 mmol) instead of 5-(bromomethyl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione and N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide, respectively.
The title compound (430 mg, yield 45%) was obtained using a method similar to the Step 2 synthesis method of Example 1 above with benzyl 4-(piperidin-4-yl)piperazin-1-carboxylate (619 mg, 2.04 mmol) instead of Boc-piperazine.
The title compound (873 mg, yield 94%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with benzyl 4-(1-(5-methoxy-2-methyl-4-nitrophenyl)piperidin-4-yl)piperazin-1-carboxylate (988 mg, 1.09 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (1.1 g, yield 74%) was obtained using a method similar to the Step 3 synthesis method of Example 35 above with benzyl 4-(1-(4-amino-5-methoxy-2-methylphenyl)piperidin-4-yl)piperazin-1-carboxylate (873 mg, 1.99 mmol) instead of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (648 mg, yield 72%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-(1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-yl)piperazin-1-carboxylate (1.1 g, 1.67 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (80 mg, yield 22%) was obtained using a method similar to the Step 1 synthesis method of Example 45 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (300 mg, 0.49 mmol) instead of N-(2-((5-chloro-2-((2-methoxy-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (49 mg, yield 74%) was obtained using a method similar to the Step 2 synthesis method of Example 45 above with N-(2-((5-chloro-2-((4-(4-(4-(2-(1,3-dioxoindolin-2-yl)ethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxy-5-methylphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (80 mg, 0.10 mmol) instead of N-(2-((5-chloro-2-((4-(4-(4-(2-(1,3-dioxoindolin-2-yl)ethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (21 mg, yield 32%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (22 mg, 0.079 mmol) and N-(2-((2-((4-(4-(4-(2-aminoethyl)piperazin-1-yl)piperidin-1-yl)-2-methoxy-5-methylphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (47 mg, 0.072 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine, respectively.
A mixture of 1-fluoro-5-methoxy-2-methyl-4-nitrobenzene (6.5 g, 35.11 mmol), benzyl 4-(piperazin-1-ylmethyl)piperidin-1-carboxylate (14.4 g, 36.86 mmol) and potassium carbonate (17.0 g, 122.87 mmol) in dimethyl sulfoxide (87 mL) was stirred at 110° C. After stirring overnight, water was added to the reaction mixture, followed by stirring, filtration, and washing with water. The remaining material was washed sequentially with ethyl acetate, followed by water and saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by column chromatography to obtain the title compound (14.0 g, 72%).
The title compound (718 mg, yield 99%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with benzyl 4-((4-(5-methoxy-2-methyl-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (794 mg, 1.60 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (1.0 g, yield 83%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with benzyl 4-((4-(4-amino-5-methoxy-2-methylphenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (718 mg, 1.58 mmol) instead of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (800 mg, yield >100%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate (1.0 g, 1.31 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (175 mg, 92%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.24 mmol) and (tertiary butoxycarbonyl)-L-alanine (54 mg, 0.29 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (148 mg, yield 98%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl (S)-(1-(4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-1-oxopropan-2-yl))carbamate (172 mg, 0.22 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (80 mg, yield 40%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (57 mg, 0.21 mmol) and N-(2-((2-((4-(4-((1-(L-alanyl)piperidin-4-yl)methyl)piperazin-1-yl)-2-methoxy-5-methylphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (145 mg, 0.21 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine, respectively.
The title compound (108 mg, yield 54%) was obtained using a method similar to the Step 2 synthesis method of Example 2 above with tertiary butyl 4-formylpiperidin-1-carboxylate (78 mg, 0.37 mmol) and N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.24 mmol) instead of 1-(3-methoxy-4-nitrophenyl)piperidin-4-one and N-Boc-piperazine, respectively.
The title compound (91 mg, yield 99%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl 4-((4-(1-(4-((5-chloro-4-((2-N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxy-5-methylphenyl)piperidin-4-yl)piperazin-1-yl)methyl)piperidin-1-carboxylate (106 mg, 0.13 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (80 mg, yield 65%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (90 mg, 0.126 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (65 mg, yield 44%) was obtained using a method similar to the Step 6 synthesis method of Example 1 above with 2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)ethyl methanesulfonate (70 mg, 0.15 mmol) and N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-piperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethylsulfonamide (93 mg, 0.15 mmol) instead of 5-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)pentanamide and N-(2-((5-chloro-2-((2-methoxy-4-(piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide.
The title compound (2.6 g, yield 97%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with benzyl 4-(piperidin-4-ylmethyl)piperazin-1-carboxylate (2.3 g, 5.79 mmol) instead of benzyl 4-(piperazin-1-ylmethyl)piperidin-1-carboxylate.
The title compound (2.38 g, yield 98%) was obtained using a method similar to the Step 2 synthesis method of Example 25 above with benzyl 4-((1-(5-methoxy-2-methyl-4-nitrophenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (2.6 g, 5.39 mmol) instead of benzyl 4-((4-(3-methoxy-4-nitrophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (4.4 g, yield 83%) was obtained using a method similar to the Step 3 synthesis method of Example 35 above with benzyl 4-((1-(4-amino-5-methoxy-2-methylphenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (3.2 g, 7.00 mmol) instead of benzyl 4-((4-(4-amino-3-chlorophenyl)piperazin-1-yl)methyl)piperidin-1-carboxylate.
The title compound (3.5 g, yield >100%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl 4-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-yl)methyl)piperazin-1-carboxylate (3.7 g, 4.85 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (183 mg, 95%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (150 mg, 0.24 mmol) and (tertiary butoxycarbonyl)-L-alanine (45 mg, 0.24 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione.
The title compound (110 mg, yield 71%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl (S)-(1-(4-((1-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-yl)methyl)piperazin-1-yl)-1-oxopropan-2-yl)carbamate (173 mg, 0.22 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (67 mg, yield 44%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (66 mg, 0.24 mmol) and N-(2-((2-((4-(4-((4-(L-alanyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-methoxy-5-methylphenyl)amino)-5-chloropyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (110 mg, 0.16 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (51 mg, yield 52%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (71 mg, 0.11 mmol) instead of 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (135 mg, 72%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (151 mg, 0.24 mmol) and (tertiary butoxycarbonyl)glycine (42 mg, 0.24 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The title compound (105 mg, yield 93%) was obtained using a method similar to the Step 5 synthesis method of Example 1 above with tertiary butyl (2-(4-((4-(4-((5-chloro-4-((2-(N-methylmethanesulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-oxoethyl)carbamate (130 mg, 0.17 mmol) instead of tertiary butyl 4-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperazin-1-carboxylate.
The title compound (65 mg, yield 48%) was obtained using a method similar to the Step 3 synthesis method of Example 33 above with 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindolin-1,3-dione (40 mg, 0.15 mmol) and N-(2-((5-chloro-2-((4-(4-((1-glycylpiperidin-4-yl)methyl)piperazin-1-yl)-2-methoxy-5-methylphenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (99 mg, 0.15 mmol) instead of 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindolin-1,3-dione and 3-(2-(benzyloxy)ethoxy)azetidine.
The title compound (105 mg, yield 70%) was obtained using a method similar to the Step 1 synthesis method of Example 6 above with 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (53 mg, 0.16 mmol) and N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (100 mg, 0.16 mmol) instead of (2R, 4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidin-2-carboxamide hydrochloride and 5-bromopentanoic acid.
The title compound (270 mg, yield 32%) was obtained using a method similar to the Step 1 synthesis method of Example 3 above with benzyl (S)-2-hydroxypropanoate (520 mg, 2.89 mmol) instead of 4-bromobutan-1-ol.
The title compound (133 mg, yield >100%) was obtained using a method similar to the Step 4 synthesis method of Example 33 above with benzyl (2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)propanoate (165 mg, 0.38 mmol) instead of 5-(3-(2-(benzyloxy)ethoxy)azetidin-1-yl)-2-(2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione.
The title compound (70 mg, 42%) was obtained using a method similar to the Step 7 synthesis method of Example 49 above with N-(2-((5-chloro-2-((2-methoxy-5-methyl-4-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)-N-methylmethanesulfonamide (109 mg, 0.17 mmol) and (2S)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)propanoic acid (60 mg, 0.17 mmol) instead of 1-((1-(4-((5-chloro-4-((2-(N-methylmethylsulfonamido)phenyl)amino)pyrimidin-2-yl)amino)-3-methoxyphenyl)piperidin-4-yl)methyl)piperidin-4-carboxylic acid hydrochloride and 3-(4-amino-1-oxoisoindolin-2-yl)piperidin-2,6-dione, respectively.
The NMR and mass results of the compounds obtained in the Examples are summarized in Table 2 below.
1H NMR (400 MHz, DMSO-d6): δ 11.13 (br s, 1H), 9.65 (br s, 1H), 8.42 (d, 1H), 8.24 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.21 (br s, 1H), 10.37 (br s, 1H), 8.74 (d, 1H), 8.28 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.13 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.12 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.12 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.13 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.11 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.12 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.23 (br s, 1H), 8.78 (d, 1H), 8.28 (m, 2H), 8.11 (s, 2H),
1H NMR (400 MHz, DMSO-d6): δ 11.05 (br s, 1H), 10.07 (br s, 1H), 8.78 (d, 1H), 8.28 (m,
1H NMR (400 MHz, DMSO-d6): δ 11.02 (br s, 1H), 9.72 (br s, 1H), 8.28 (m, 2H), 8.11 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.03 (br s, 1H), 9.73 (br s, 1H), 8.28 (m, 2H), 8.12 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 9.00 (s, 1H), 8.63 (t, 1H), 8.28 (m, 2H), 8.13 (s, 1H), 8.10
1H NMR (400 MHz, DMSO-d6): δ 8.99 (br s, 1H), 8.62 (br t, 1H), 8.29 (m, 1H), 8.28 br s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 9.47 (s, 1H), 8.99 (s, 1H), 8.62 (t, 1H), 8.39 (s, 1H), 7.27
1H NMR (400 MHz, DMSO-d6): δ 11.06 (br s, 1H), 9.45 (br s, 1H), 8.39 (br s, 1H), 8.27 (m,
1H NMR (400 MHz, DMSO-d6): δ 11.11 (br s, 1H), 9.43 (br s, 1H), 8.38 (br s, 1H), 8.26 (m,
1H NMR (400 MHz, DMSO-d6): δ 11.12 (br s, 1H), 9.47 (br s, 1H), 8.39 (br s, 1H), 8.27 (m,
1H NMR (400 MHz, DMSO-d6): δ 10.99 (br s, 1H), 10.06 (br s, 1H), 8.28 (m, 2H), 8.12 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.01 (br s, 1H), 9.72 (br s, 1H), 9.45 (br s, 1H), 8.39 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.01 (br s, 1H), 9.74 (br s, 1H), 8.28 (m, 2H), 8.09 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.07 (br s, 1H), 8.28 (m, 2H), 8.10 (s, 1H), 8.09 (br s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.11 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.11 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.13 (br s, 1H), 8.29 (br s, 1H), 8.28 (m, 1H), 8.14 (br s,
1H NMR (400 MHz, DMSO-d6): δ 9.00 (s, 1H), 8.52 (t, 1H), 8.28 (m, 2H), 8.14 (s, 1H), 8.10
1H NMR (400 MHz, DMSO-d6): δ 9.00 (s, 1H), 8.58 (t, 1H), 8.28 (m, 2H), 8.14 (s, 1H), 8.10
1H NMR (400 MHz, DMSO-d6): δ 11.12 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.14 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.08 (br s, 1H), 8.28 (br s, 1H), 8.27 (m, 1H), 8.13 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.09 (br s, 1H), 8.28 (m, 2H), 8.14 (brs, 1H), 8.10 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.09 (br s, 1H), 8.28 (m, 2H), 8.13 (brs, 1H), 8.10 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.08 (br s, 1H), 8.28 (m, 2H), 8.14 (brs, 1H), 8.10 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.18 (br s, 1H), 8.29 (br s, 1H), 8.28 (m, 1H), 8.13 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.09 (br s, H), 8.29 (br s, 1H), 8.28 (m, 1H), 8.15 (br s,
1H NMR (400 MHz, DMSO-d6): δ 11.07 (br s, 1H), 8.28 (m, 2H), 8.10 (s, 2H), 7.68 (d, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.07 (br s, 1H), 8.28 (m, 2H), 8.10 (s, 2H), 7.65 (d, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.15 (br s, 1H), 8.28 (m, 2H), 8.14 (br s, 1H), 8.11 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.09 (br s, 1H), 8.33 (br s, 1H), 8.22 (m, 1H), 8.14 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.09 (br s, 1H), 8.32 (br s, 1H), 8.23 (m, 1H), 8.14 (s, 1H),
1H NMR (400 MHz, DMSO-d6): δ 11.11 (br s, 1H), 8.32 (br s, 1H), 8.22 (m, 1H), 8.13 (s, 1H),
Evaluation of the Compounds of the Present Invention
The ability of the compounds of the present invention to degrade the mutant EGFR protein was confirmed in Ba/F3 cells expressing normal EGFR, mutant EGFRdel19+T790M+C797S or mutant EGFRL858R+T790M+C797S. A Western blot test using the EGFR antibody was performed.
Specifically, Ba/F3 cells expressing EGFR were cultured in RPMI 1640 (Gibco, NY, USA) containing 10% fetal bovine serum (FBS, Gibco, NY, USA). Cells expressing normal EGFR, mutant EGFRdel19+T790M+C797S or mutant EGFRL858R+T790M+C797S were treated with the compounds of the present invention at concentrations of 0.1 μM and 1 μM, respectively, for 24 hours. After that, it was washed twice with phosphate buffered saline (PBS) and SDS-PAGE was performed with 30 μg of cell lysate. After the protein separated on the gel was transferred to a nitrocellulose membrane, the expression level of the EGFR protein was confirmed using an EGFR antibody (Santa Cruz biotechnology, Santacruz, CA, USA). EGFR bands separated by Western blotting were quantitatively analyzed with Fluo-S MultiImager (Bio-Rad, Hercules, CA, USA).
The experimental results are shown in Table 3 below.
Pharmacokinetic tests for the compounds of the present invention were conducted as below. After a single oral administration of the compound of the present invention to ICR mice, the concentration of the compound over time was followed up and analyzed.
The compound of the present invention was suspended in 10% DMSO/90% (30% HPbCD aqueous solution) and orally administered to mice at a dose of 5 mg/kg. Blood was collected at a fixed time and plasma was separated. Drug analysis was performed using HPLC (XBridge column C18, Waters, mobile phase 0.1% formic acid:acetonitrile (30:70, %/%)) and MS/MS (ESI positive, MRM). Mouse blood plasma and each commercially available standard solution were mixed at a ratio of 9:1 to prepare and calibrate at concentrations of 5, 50, 100, 500, 100 and 5,000 ng/mL. In addition, the preparation of QC samples was prepared by mixing the mouse blood plasma and the standard solution for QC at a ratio of 9:1, and prepared to be concentrations of 100, 750 and 2,500 ng/mL. 100 μL of the plasma sample was transferred to a centrifugation tube, and 10 μL of the internal standard solution and 300 μL of methanol were added, followed by mixing for about 30 seconds for pretreatment. The tube was centrifuged at 3,000×g (4° C.) for about 5 minutes, and the supernatant was taken and transferred to an LC vial and injected into the instrument. Then, the concentration of the compound in mouse plasma was quantified by applying a previously validated assay. For pharmacokinetic parameters, the program WinNonlin 5.2 (Pharsight, USA) was used. AUC0-t, AUC0-∞, Cmax, Tmax, and t1/2 were calculated by Noncompartment modeling (best fit). The pharmacokinetic parameter results were expressed as the mean (Mean) and standard deviation (SD), and were statistically processed using the SPSS program (Statistical Package for the Social Sciences, 10.0K, USA).
The test results are shown in Table 4 below.
All documents mentioned herein are incorporated herein by reference as if their contents were set forth herein. When introducing elements of the present invention or preferred embodiment(s) thereof, the articles “a”, “an”, “the” and “said” refer to one or more elements is intended to mean that there is. The terms “comprising” ˜, “including” and “having” are intended to be inclusive and mean that there may be additional elements other than those listed. Although the invention has been described in terms of a particular embodiment or embodiments, it should not be construed as limiting the details of these embodiments.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2020-0117186 | Sep 2020 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2021/011898 | 9/2/2021 | WO |
